U.S. patent application number 15/502009 was filed with the patent office on 2017-08-10 for methods for treating inflammatory bowel disease using prostate specific membrane antigen (psma) inhibitors.
This patent application is currently assigned to THE JOHNS HOPKINS UNIVERSITY. The applicant listed for this patent is THE JOHNS HOPKINS UNIVERSITY. Invention is credited to XUHANG LI, RANA RAIS, BARBARA SLUSHER.
Application Number | 20170224709 15/502009 |
Document ID | / |
Family ID | 55264768 |
Filed Date | 2017-08-10 |
United States Patent
Application |
20170224709 |
Kind Code |
A1 |
SLUSHER; BARBARA ; et
al. |
August 10, 2017 |
METHODS FOR TREATING INFLAMMATORY BOWEL DISEASE USING PROSTATE
SPECIFIC MEMBRANE ANTIGEN (PSMA) INHIBITORS
Abstract
Methods and compounds are disclosed for treating inflammatory
bowel disease (IBD) by using Prostate Specific Membrane Antigen
(PSMA) inhibitors.
Inventors: |
SLUSHER; BARBARA;
(KINGSVILLE, MD) ; RAIS; RANA; (WEST FIRENDSHIP,
MD) ; LI; XUHANG; (CLARKSVILLE, MD) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
THE JOHNS HOPKINS UNIVERSITY |
BALTIMORE |
MD |
US |
|
|
Assignee: |
THE JOHNS HOPKINS
UNIVERSITY
BALTIMORE
MD
|
Family ID: |
55264768 |
Appl. No.: |
15/502009 |
Filed: |
August 6, 2015 |
PCT Filed: |
August 6, 2015 |
PCT NO: |
PCT/US15/44025 |
371 Date: |
February 6, 2017 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
62033948 |
Aug 6, 2014 |
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 1/00 20180101; A61P
29/00 20180101; A61K 31/662 20130101; A61K 9/0053 20130101; A61K
9/0031 20130101; A61K 9/02 20130101; A61K 31/194 20130101 |
International
Class: |
A61K 31/662 20060101
A61K031/662; A61K 9/00 20060101 A61K009/00; A61K 9/02 20060101
A61K009/02; A61K 31/194 20060101 A61K031/194 |
Goverment Interests
FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT
[0002] This invention was made with government support under R01
CA161056-01 awarded by the National Cancer Institute (NCI). The
government has certain rights in the invention.
Claims
1. A method for treating an inflammatory bowel disease (IBD) in a
subject in need thereof, the method comprising administering to the
subject a therapeutically effective amount of a Prostate Specific
Membrane Antigen (PSMA) inhibitor.
2. The method of claim 1, wherein the PSMA inhibitor is
2-(phosphonomethyl)-pentanedioic acid (2-PMPA), having the
structure: ##STR00010##
3. The method of claim 2, wherein the IBD is selected from the
group consisting of Crohn's disease (CD) and ulcerative colitis
(UC).
4. The method of claim 3, wherein the 2-PMPA is administered to the
subject parenterally.
5. The method of claim 1, wherein the PSMA inhibitor is
(3-2-Mercaptoethyl)biphenyl-2,3-dicarboxylic acid (E2072), having
the structure: ##STR00011##
6. The method of claim 5, wherein the IBD is selected from the
group consisting of Crohn's disease (CD) and ulcerative colitis
(UC).
7. The method of claim 6, wherein the E2072 is administered to the
subject parenterally.
8. The method of claim 1, wherein the PSMA inhibitor is an ester
prodrug of 2-(phosphonomethyl)-pentanedioic acid (2-PMPA), wherein
acidic moieties in the ester prodrug have been capped with
pivaloyloxymethyl (POM) or propyloxycarbonyloxymethyl (POC).
9. The method of claim 8, wherein the ester prodrug of 2-PMPA is a
phosphonate or carboxylate prodrug of 2-PMPA.
10. The method of claim 9, wherein the ester prodrug of 2-PMPA is
Tris-propyloxycarbonyloxymethyl-2-(phosphonomethyl)-pentanedioic
acid (Tris-POC-2-PMPA), having the structure: ##STR00012##
11. The method of claim 10, wherein the IBD is selected from the
group consisting of Crohn's disease (CD) and ulcerative colitis
(UC).
12. The method of claim 11, wherein the ester prodrug of 2-PMPA is
administered to the subject orally.
13. The method of claim 11, wherein the Tris-POC-2-PMPA is
administered to the subject orally.
14. The method of claim 11, wherein the ester prodrug or 2-PMPA is
administered to the subject via a suppository.
15. The method of claim 14, wherein the suppository is a rectal
suppository.
16. The method of claim 11, wherein the Tris-POC-2-PMPA is
administered to the subject via a suppository.
17. The method of claim 16, wherein the suppository is a rectal
suppository.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional
Application No. 62/033,948, filed Aug. 6, 2014, which is
incorporated herein by reference in its entirety.
BACKGROUND
[0003] Inflammatory Bowel Disease (IBD), an idiopathic, chronic and
frequently disabling inflammatory disorder of the intestine, has
two subtypes: Crohn's disease (CD) and ulcerative colitis (UC),
each accounting for approximately 50% of IBD patients (Xavier and
Podolsky, 2007; Srober et al., 2007; Sartor, 2006). IBD is a
widespread GI disease, with a prevalence of approximately 0.2% in
the Western population. In the United States alone, there are 1.4
million diagnosed IBD patients, resulting in enormous suffering and
health-care costs. It is increasingly clear that IBD is a complex
multifactorial disease with both genetic and environmental
contributions, the interaction of which leads to IBD (Xavier and
Podolsky; Strober et al., 2007; Sartor, 2006; Kaser et al., 2010).
Unfortunately, the etiology of this mucosal dysregulation in UC and
CD remain elusive (Kaser et al., 2010). Despite increasing
therapeutic options available for the management of IBD,
approximately 1/3 of IBD patients do not respond to any given
therapy, and there is no cure for IBD (Hamilton et al., 2012).
Anti-tumor necrosis factor (TNF)-based therapies, such as
infliximab (IFX), adalimumab and certolizumab pegol are currently
the most effective therapies for severe UC and CD (Hanauer et al.,
2002; Kozuch and Hanauer, 2008; Colombel et al., 2007; Schreiber et
al., 2007). However, one-third of patients with CD do not respond
to anti-TNF therapies and another third lose responsiveness within
six months of initiating therapy (Regueiro et al., 2007; Lawrance,
2014). These non-responders have more aggressive mucosal immune
responses and additional treatments are indicated (Schmidt et al.,
2007). Patients with extensive disease or who are at risk for short
gut syndrome due to prior resections are usually poor surgical
candidates. Currently, the only approved medication for patients
who have failed an anti-TNF agent is natalizumab. However,
natalizumab has been associated with several cases of progressive
and often fatal multifocal leukoencephalopathy (PML) (Van et al.,
2005). This emphasizes the significance of exploring and
identifying new and more effective therapies in patients with
IBD.
SUMMARY
[0004] In one aspect, the presently disclosed subject matter
provides a method for treating an inflammatory bowel disease (IBD)
in a subject in need thereof, the method comprising administering
to the subject a therapeutically effective amount of a Prostate
Specific Membrane Antigen (PSMA) inhibitor.
[0005] In a particular aspect, the PSMA inhibitor for use in the
methods of the presently disclosed subject matter is
2-(phosphonomethyl)-pentanedioic acid (2-PMPA), having the
structure:
##STR00001##
In further aspects, the IBD is selected from the group consisting
of Crohn's disease (CD) and ulcerative colitis (UC), and the 2-PMPA
is administered parenterally.
[0006] In another particular aspect, the PSMA inhibitor for use in
the methods of the presently disclosed subject matter is
(3-2-Mercaptoethyl)biphenyl-2,3-dicarboxylic acid (E2072), having
the structure:
##STR00002##
In further aspects, the IBD is selected from the group consisting
of CD and UC, and the E2072 is administered parenterally.
[0007] In another particular aspect, the PSMA inhibitor for use in
the methods of the presently disclosed subject matter is an ester
prodrug of 2-PMPA (e.g. a phosphonate or carboxylate prodrug of
2-PMPA), wherein acidic moieties in the ester prodrug have been
capped with pivaloyloxymethyl (POM) or propyloxycarbonyloxymethyl
(POC). In yet another particular aspect, the ester prodrug of
2-PMPA is
Tris-propyloxycarbonyloxymethyl-2-(phosphonomethyl)-pentanedioic
acid (Tris-POC-2-PMPA), having the structure:
##STR00003##
[0008] In further aspects, the IBD is selected from the group
consisting of CD and UC, and the ester prodrug of 2-PMPA is
administered orally. In further aspects, the IBD is selected from
the group consisting of CD and UC, and the Tris-POC-2-PMPA is
administered orally. In further aspects, the IBD is selected from
the group consisting of CD and UC, and the ester prodrug of 2-PMPA
is administered via a suppository. In further aspects, the IBD is
selected from the group consisting of CD and UC, and the
Tris-POC-2-PMPA is administered via a suppository. In some
embodiments, the suppository is a rectal suppository.
[0009] Certain aspects of the presently disclosed subject matter
having been stated hereinabove, which are addressed in whole or in
part by the presently disclosed subject matter, other aspects will
become evident as the description proceeds when taken in connection
with the accompanying Examples and Figures as best described herein
below.
BRIEF DESCRIPTION OF THE FIGURES
[0010] Having thus described the presently disclosed subject matter
in general terms, reference will now be made to the accompanying
Figures, which are not necessarily drawn to scale, and wherein:
[0011] FIG. 1A and FIG. 1B show the marked increase of PSMA
expression in the villous epithelium from ileal sample of CD
patient (Zhang et al., 2012). Immunohistochemical localization of
PSMA (indicated by arrows) in: (FIG. 1A) control non-IBD subject;
and (FIG. 1B) diseased ileal mucosa from the proximal margin of
resected ileum from an ileal CD subject. Magnification is
100.times.. Bar is 200 mm;
[0012] FIG. 2A and FIG. 2B show the marked elevation of PMSA
activity in the inflamed (disease) intestinal mucosa of patients
with IBD. PMSA activity was measured from mucosa specimens from
involved (inflamed with active disease) and uninvolved
(macroscopically normal, as a control) from IBD patients or from
non-IBD controls (healthy controls or patients with diverticulitis)
(n=20): (FIG. 2A) comparison between IBD (active disease vs.
normal/uninvolved tissues) vs. non-IBD controls; and (FIG. 2B)
comparison between active disease vs. normal/uninvolved of the same
patients (CD, Crohn's disease; UC, ulcerative colitis; the Arabic
numbers refer to different patients). Note: GCPII is also highly
upregulated in colon cancer (see A). *P<0.05;
[0013] FIG. 3 shows that PSMAi (2-PMPA) ameliorates disease
activity in DSS-induced murine model of colitis. C57/B6 mice
(approximately 8 weeks old) that were induced to develop colitis
with DSS (2.5%, 7 days in drinking water) were treated
simultaneously with the vehicle or 2-PMPA (100 mg/kg),
respectively. Disease activity index (DAI), which positively
correlated with the disease severity, was used as a measure for
clinical activity. *P<0.05;
[0014] FIG. 4 shows that PSMAi (2-PMPA) effectively suppresses PSMA
activity in the colonic or cecal mucosa of DSS-induced murine model
of colitis. PSMA activity was measured using extract from
mucosa;
[0015] FIG. 5 shows that 2-PMPA treatment reduces size and weight
of spleen DSS-induced murine model of colitis;
[0016] FIG. 6A, FIG. 6B, FIG. 6C, and FIG. 6D show that PSMAi
(2-PMPA) treatment leads to not only improvement of disease but
even retraction of prolapse in IL-10 knockout (IL-10 KO) mice that
spontaneously develop colitis. IL-10 KO mice (C57/B6; 3 month old)
were treated with 2-PMPA (100 mg/kg) for 2 weeks: (FIG. 6A)
improvement of prolapse and colonic macroscopic disease
(inflammation, hypertrophy, stool inconsistency); (FIG. 6B) colon
weight changes; (FIG. 6C) body weight after 2-PMPA; and (FIG. 6D)
prolapse retraction after treatment. *P<0.05;
[0017] FIG. 7 shows that E2072, a PSMAi that is structurally
distinct from 2-PMPA, also ameliorates disease activity in
DSS-induced murine model of colitis. Experiments were performed as
described in FIG. 3. Disease activity index (DAI), which positively
correlated with the disease severity, was used as a measure for
clinical activity. *P<0.05;
[0018] FIG. 8A and FIG. 8B show orally available Tris POC 2-PMPA
prodrug: (FIG. 8A) plasma 2-PMPA concentrations following 30 mg/kg
per oral administration of 2-PMPA (red squares) or 2-PMPA prodrug
(black circles). 2-PMPA prodrug achieved 20-30-fold enhancements in
permeability; and (FIG. 8B) comparison of plasma and colonic 2-PMPA
concentrations following 100 mg/kg i.p. administration of 2-PMPA
(red bars) or 2-PMPA prodrug (blue bars). 2-PMPA prodrug achieved
concentrations similar to 2-PMPA i.p. dose which was efficacious in
both DSS and IL-10 knockout models; and
[0019] FIG. 9 shows that prodrug Tris POC 2-PMPA treatment reverses
the colon shortening of DSS colitis mice. The p value (P-0.06) was
close but did not reach statistic significance, at least in part
due to the small number of mice in each group (n=5 per group).
[0020] The patent or application file contains at least one drawing
executed in color. Copies of this patent or patent application
publication with color drawings will be provided by the Office upon
request and payment of the necessary fee.
DETAILED DESCRIPTION
[0021] The presently disclosed subject matter now will be described
more fully hereinafter with reference to the accompanying Figures,
in which some, but not all embodiments of the inventions are shown.
Like numbers refer to like elements throughout. The presently
disclosed subject matter may be embodied in many different forms
and should not be construed as limited to the embodiments set forth
herein; rather, these embodiments are provided so that this
disclosure will satisfy applicable legal requirements. Indeed, many
modifications and other embodiments of the presently disclosed
subject matter set forth herein will come to mind to one skilled in
the art to which the presently disclosed subject matter pertains
having the benefit of the teachings presented in the foregoing
descriptions and the associated Figures. Therefore, it is to be
understood that the presently disclosed subject matter is not to be
limited to the specific embodiments disclosed and that
modifications and other embodiments are intended to be included
within the scope of the appended claims.
[0022] The practice of the present invention will typically employ,
unless otherwise indicated, conventional techniques of cell
biology, cell culture, molecular biology, transgenic biology,
microbiology, recombinant nucleic acid (e.g., DNA) technology,
immunology, and RNA interference (RNAi) which are within the skill
of the art. Non-limiting descriptions of certain of these
techniques are found in the following publications: Ausubel, F., et
al., (eds.), Current Protocols in Molecular Biology, Current
Protocols in Immunology, Current Protocols in Protein Science, and
Current Protocols in Cell Biology, all John Wiley & Sons, N.Y.,
edition as of December 2008; Sambrook, Russell, and Sambrook,
Molecular Cloning. A Laboratory Manual, 3.sup.rd ed., Cold Spring
Harbor Laboratory Press, Cold Spring Harbor, 2001; Harlow, E. and
Lane, D., Antibodies--A Laboratory Manual, Cold Spring Harbor
Laboratory Press, Cold Spring Harbor, 1988; Freshney, R. I.,
"Culture of Animal Cells, A Manual of Basic Technique", 5th ed.,
John Wiley & Sons, Hoboken, N.J., 2005. Non-limiting
information regarding therapeutic agents and human diseases is
found in Goodman and Gilman's The Pharmacological Basis of
Therapeutics, 11th Ed., McGraw Hill, 2005, Katzung, B. (ed.) Basic
and Clinical Pharmacology, McGraw-Hill/Appleton & Lange
10.sup.th ed. (2006) or 11th edition (July 2009). Non-limiting
information regarding genes and genetic disorders is found in
McKusick, V. A.: Mendelian Inheritance in Man. A Catalog of Human
Genes and Genetic Disorders. Baltimore: Johns Hopkins University
Press, 1998 (12th edition) or the more recent online database:
Online Mendelian Inheritance in Man, OMIM.TM.. McKusick-Nathans
Institute of Genetic Medicine, Johns Hopkins University (Baltimore,
Md.) and National Center for Biotechnology Information, National
Library of Medicine (Bethesda, Md.), as of May 1, 2010, World Wide
Web URL: http://www.ncbi.nlm.nih.gov/omim/ and in Online Mendelian
Inheritance in Animals (OMIA), a database of genes, inherited
disorders and traits in animal species (other than human and
mouse), at http://omia.angis.org.au/contact.shtml.
[0023] Immune-mediated gastrointestinal disorders encompass a wide
range of debilitating gastrointestinal diseases of various
etiologies. One such immune-mediated gastrointestinal disorder,
inflammatory bowel disease (IBD), is the collective term used to
describe two gastrointestinal disorders of unknown etiology:
Crohn's disease (CD) and ulcerative colitis (UC). The course and
prognosis of IBD, which occurs world-wide and is reported to
afflict as many as two million people, varies widely. The onset of
IBD typically occurs during young adulthood, with the most common
symptoms being diarrhea, abdominal pain, and fever. The diarrhea
may range from mild to severe and in ulcerative colitis often is
accompanied by bleeding. Anemia and weight loss are additional
common signs of IBD. Ten percent to fifteen percent of all patients
with IBD will require surgery over a ten year period. In addition,
patients with IBD are at increased risk for the development of
intestinal cancer. Increased occurrence of psychological problems,
including anxiety and depression, are perhaps not surprising
symptoms of what is often a debilitating disease that strikes
people in the prime of life.
[0024] 6-Mercaptopurine (6-MP) and azathioprine (AZA), a pro-drug
that is non-enzymatically converted to 6-mercaptopurine (6-MP), are
6-MP drugs that have been used as treatments for inflammatory bowel
diseases such as Crohn's disease and ulcerative colitis (Kirschner,
1998). 6-MP can be enzymatically converted to various 6-MP
metabolites, including 6-methyl-mercaptopurine (6-MMP) and
6-thioguanine (6-TG) and their nucleotides. 6-TG nucleotides are
thought to be the active metabolite in mediating many of the
effects of 6-MP drug treatment.
[0025] Thiopurine methyltransferase (TPMT) is a cytoplasmic enzyme
that preferentially catalyzes the S-methylation of 6-MP and 6-TG to
form S-methylated metabolites such as 6-MMP and 6-methylthioguanine
(6-MTG), respectively. TPMT exhibits genetic polymorphism, with 89%
of Caucasians and African Americans having high activity, 11%
intermediate activity and 1 in 300 TPMT deficient. Clinical studies
with AZA and 6-MP have shown an inverse relationship between TPMT
activity and 6-TGN accumulation. Patients who less efficiently
methylate these thiopurines have more extensive conversion to
6-TGN, which can lead to potentially fatal hematopoietic toxicity.
Therefore, patients who have less active TPMT can be more
susceptible to toxic side effects of 6-MP therapy.
[0026] Although drugs such as 6-MP and AZA have been used for
treating IBD, non-responsiveness and drug toxicity unfortunately
have complicated treatment in some patients. Complications
associated with 6-MP drug treatment include allergic reactions,
neoplasia, opportunistic infections, hepatitis, bone marrow
suppression, and pancreatitis. Therefore, many physicians are
reluctant to treat patients with AZA because of its potential side
effects, especially infection and neoplasia.
[0027] Anti-tumor necrosis factor (TNF)-based therapies, such as
infliximab (IFX), adalimumab and certolizumab pegol are currently
the most effective therapies for severe UC and CD (Hanauer et al.,
2002; Kozuch and Hanauer, 2008; Colombel et al., 2007; Schreiber et
al., 2007). Despite increasing therapeutic options available for
the management of IBD, approximately 1/3 of IBD patients do not
respond to any given therapy, and there is no cure for IBD
(Hamilton et al., 2012). For example, one-third of patients with CD
do not respond to anti-TNF therapies and another third lose
responsiveness within six months of initiating therapy (Regueiro et
al., 2007; Lawrance, 2014). These non-responders have more
aggressive mucosal immune responses and additional treatments are
indicated (Schmidt et al., 2007). Patients with extensive disease
or who are at risk for short gut syndrome due to prior resections
are usually poor surgical candidates. Currently, the only approved
medication for patients who have failed an anti-TNF agent is
natalizumab. However, natalizumab has been associated with several
cases of progressive and often fatal multifocal leukoencephalopathy
(PML) (Van et al., 2005). This emphasizes the significance of
exploring and identifying new and more effective therapies in
patients with IBD.
[0028] As described more fully in the Examples below, the presently
disclosed subject matter relates to the discovery that Prostate
Specific Membrane Antigen (PSMA) enzymatic activity is consistently
and robustly activated in human IBD, and that pharmacological
inhibition of PSMA using multiple structurally distinct inhibitors
ameliorate IBD symptoms in two murine preclinical models.
I. Methods of Treating Inflammatory Bowel Disease with a Prostate
Specific Membrane Antigen Inhibitor
[0029] In one embodiment, the presently disclosed subject matter
provides a method for treating Inflammatory Bowel Disease (IBD) in
a subject in need thereof with a therapeutically effective amount
of a Prostate Specific Membrane Antigen (PSMA) inhibitor.
[0030] PSMA, also termed Glutamate Carboxypeptidase II (GCPII) and
Folate Hydrolase I (FOLH1), is a metallopeptidase that catalyzes
the hydrolysis of N-acetylated aspartate-glutamate (NAAG) to
N-acetyl aspartate (NAA) and glutamate and cleaves terminal
glutamate moieties sequentially from folate polyglutamate (Ristau
et al., 2013; Mesters et al., 2006; Slusher et al., 2013). As used
herein, the terms "PSMA" or "PSMA polypeptide" refer to a naturally
occurring or endogenous PSMA and to proteins having an amino acid
sequence which is the same as that of a naturally occurring or
endogenous PSMA (e.g., recombinant proteins). Accordingly, as
defined herein, the term includes mature PSMA, glycosylated or
unglycosylated PSMA proteins, polymorphic or allelic variants, and
other isoforms of PSMA (e.g., produced by alternative splicing or
other cellular processes).
[0031] As used herein, a PSMA inhibitor is a molecule that
decreases or inhibits the activity of PSMA. The modulation of the
activity of PSMA may be detected by use of an assay for the
intrinsic N-acetylated alpha-linked acidic dipeptidase (NAALADase)
activity of PSMA (Tang et al., 2003; Robinson et al. 1987; Lupold
et al., 2002; U.S. Patent App. Pub. No. 20110064657). Inhibition
curves may be determined using semi-log plots and IC.sub.50 values
determined at the concentration at which enzyme activity was
inhibited by 50%. In some embodiments, the PSMA inhibitor has an
IC.sub.50 value ranging from about 0.1 to about 200 nM. In a
further embodiment, the PSMA inhibitor has an IC.sub.50 value
ranging from about 0.5 to about 118 nM.
[0032] The PSMA inhibitor may interact with PSMA directly (e.g.,
via interaction with the binding site of PSMA) or may interact with
another molecule that results in a decrease in the activity of
PSMA. The binding site of PSMA contains a binuclear zinc ion and
two substrate binding pockets, i.e., an S1 (nonpharmacophore)
pocket and an (pharmacophore) pocket. The active site also contains
a chloride ion in the S1 pocket. In the vicinity of the S1 pocket
resides a funnel-shaped tunnel with a depth of approximately 20
.ANG. and a width of 8-9 .ANG.. Similarly, a narrow cavity is
present near the S1' pocket.
[0033] In one embodiment, the PSMA inhibitor for use within the
methods of the presently disclosed subject matter is
2-(phosphonomethyl)-pentanedioic acid (2-PMPA), having the
structure:
##STR00004##
[0034] In another embodiment, the PSMA inhibitor for use within the
methods of the presently disclosed subject matter is
(3-2-Mercaptoethyl)biphenyl-2,3-dicarboxylic acid (E2072), having
the structure:
##STR00005##
[0035] In another embodiment, the PSMA inhibitor for use within the
methods of the presently disclosed subject matter is an ester
prodrug of 2-PMPA (e.g. a phosphonate or carboxylate prodrug of
2-PMPA) wherein acidic moieties have been capped with
pivaloyloxymethyl (POM) or propyloxycarbonyloxymethyl (POC).
Non-limiting examples of ester prodrugs of 2-PMPA of use herein are
described in international PCT Application entitled "Prodrugs of
PSMA Inhibitor" filed concurrently herewith (Attorney Docket No.:
11232-00435.P13008-02), which is herein incorporated by reference
in its entirety. In one embodiment, the ester prodrug of 2-PMPA is
Tris-propyloxycarbonyloxymethyl-2-(phosphonomethyl)-pentanedioic
acid (Tris-POC-2-PMPA), having the structure:
##STR00006##
[0036] As described more fully in the Examples below, there is a
marked elevation of PMSA activity in the diseased intestinal mucosa
of subjects with IBD. As used herein, the term "elevated PSMA
activity" means an increase of PSMA activity in a subject with IBD
as compared to the PSMA activity in a subject without IBD, such as
an increase of approximately 100%, 200%, 300%, 400%, 500%, 600%,
700%, 800%, 900%, 1000%, or more.
[0037] In some embodiments, the presently disclosed subject matter
provides methods for inhibiting PMSA activity in a subject with
IBD. As used herein, the term "inhibit" means to decrease or
diminish PSMA activity in a subject in need thereof. The term
"inhibit" also may mean to decrease, suppress, attenuate, diminish,
arrest, or stabilize the development or progression of a disease or
condition, such as IBD. Inhibition may occur, for e.g., by at least
10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or even
100% compared to an untreated control subject or a subject without
the disease or disorder.
[0038] In general, using the presently disclosed methods to treat
the IBD in a subject results in a decrease in the severity of the
IBD. As used herein, the terms "treat," "treating," "treatment,"
and the like refer to reducing or ameliorating a disease or
condition, such as IBD, and/or symptoms associated therewith. It
will be appreciated that, although not precluded, treating a
disease or condition, such as IBD, does not require that the
disorder, condition or symptoms associated therewith be completely
eliminated. The term "decrease" is meant to inhibit, suppress,
attenuate, diminish, arrest, or stabilize at least one symptom of
IBD (e.g., rectal prolapse, gut inflammation, colonic hypertrophy,
stool inconsistency, and the like).
[0039] IBD has been classified into the broad categories of Crohn's
disease and ulcerative colitis. Accordingly, as used herein, "a
subject having inflammatory bowel disease" is synonymous with the
term "a subject diagnosed with having an inflammatory bowel
disease," and means a patient having Crohn's disease or ulcerative
colitis. Crohn's disease (regional enteritis) is a disease of
chronic inflammation that can involve any part of the
gastrointestinal tract. Commonly, the distal portion of the small
intestine (ileum) and cecum are affected. In other cases, the
disease is confined to the small intestine, colon or anorectal
region. Crohn's disease occasionally involves the duodenum and
stomach, and more rarely the esophagus and oral cavity.
[0040] The variable clinical manifestations of Crohn's disease are,
in part, a result of the varying anatomic localization of the
disease. The most frequent symptoms of CD are abdominal pain,
diarrhea and recurrent fever. CD is commonly associated with
intestinal obstruction or fistula, which is an abnormal passage
between diseased loops of bowel, for example. Crohn's disease also
includes complications such as inflammation of the eye, joints and
skin; liver disease; kidney stones or amyloidosis. In addition, CD
is associated with an increased risk of intestinal cancer.
[0041] Several features are characteristic of the pathology of
Crohn's disease. The inflammation associated with CD, known as
transmural inflammation, involves all layers of the bowel wall.
Thickening and edema, for example, typically also appear throughout
the bowel wall, with fibrosis also present in long-standing
disease. The inflammation characteristic of CD also is
discontinuous in that segments of inflamed tissue, known as "skip
lesions," are separated by apparently normal intestine.
Furthermore, linear ulcerations, edema, and inflammation of the
intervening tissue lead to a "cobblestone" appearance of the
intestinal mucosa, which is distinctive of CD.
[0042] A hallmark of Crohn's disease is the presence of discrete
aggregations of inflammatory cells, known as granulomas, which are
generally found in the submucosa. Some Crohn's disease cases
display the typical discrete granulomas, while others show
nonspecific transmural inflammation. As a result, the presence of
discrete granulomas is indicative of CD, although the absence of
granulomas also is consistent with the disease. Thus, transmural or
discontinuous inflammation, rather than the presence of granulomas,
is a preferred diagnostic indicator of Crohn's disease (Rubin and
Farber, 1994).
[0043] Ulcerative colitis (UC) is a disease of the large intestine
characterized by chronic diarrhea with cramping abdominal pain,
rectal bleeding, and loose discharges of blood, pus and mucus. The
manifestations of ulcerative colitis vary widely. A pattern of
exacerbations and remissions typifies the clinical course of most
UC patients (70%), although continuous symptoms without remission
are present in some patients with UC. Local and systemic
complications of UC include arthritis, eye inflammation such as
uveitis, skin ulcers and liver disease. In addition, ulcerative
colitis and especially long-standing, extensive disease is
associated with an increased risk of colon carcinoma.
[0044] Several pathologic features characterize UC in distinction
to other inflammatory bowel diseases. Ulcerative colitis is a
diffuse disease that usually extends from the most distal part of
the rectum for a variable distance proximally. The term left-sided
colitis describes an inflammation that involves the distal portion
of the colon, extending as far as the splenic flexure. Sparing of
the rectum or involvement of the right side (proximal portion) of
the colon alone is unusual in ulcerative colitis. The inflammatory
process of ulcerative colitis is limited to the colon and does not
involve, for example, the small intestine, stomach or esophagus. In
addition, ulcerative colitis is distinguished by a superficial
inflammation of the mucosa that generally spares the deeper layers
of the bowel wall. Crypt abscesses, in which degenerated intestinal
crypts are filled with neutrophils, also are typical of ulcerative
colitis (Rubin and Farber, 1994).
[0045] In comparison with Crohn's disease, which is a patchy
disease with frequent sparing of the rectum, ulcerative colitis is
characterized by a continuous inflammation of the colon that
usually is more severe distally than proximally. The inflammation
in ulcerative colitis is superficial in that it is usually limited
to the mucosal layer and is characterized by an acute inflammatory
infiltrate with neutrophils and crypt abscesses. In contrast,
Crohn's disease affects the entire thickness of the bowel wall with
granulomas often, although not always, present. Disease that
terminates at the ileocecal valve, or in the colon distal to it, is
indicative of ulcerative colitis, while involvement of the terminal
ileum, a cobblestone-like appearance, discrete ulcers or fistulas
suggest Crohn's disease.
II. Pharmaceutical Compositions and Administration
[0046] In another aspect, the present disclosure provides a
pharmaceutical composition including a PSMA inhibitor alone or in
combination with one or more additional therapeutic agents in
admixture with a pharmaceutically acceptable excipient. One of
skill in the art will recognize that the pharmaceutical
compositions include the pharmaceutically acceptable salts of the
compounds described above.
[0047] In therapeutic and/or diagnostic applications, the PSMA
inhibitor for use within the methods of the presently disclosed
subject matter can be formulated for a variety of modes of
administration, including oral, systemic, and topical or localized
administration. Techniques and formulations generally may be found
in Remington: The Science and Practice of Pharmacy (20.sup.th ed.)
Lippincott, Williams & Wilkins (2000).
[0048] In one embodiment, where the PSMA inhibitor for use within
the methods of the presently disclosed subject matter is an ester
prodrug of 2-PMPA wherein acidic moieties have been capped with
pivaloyloxymethyl (POM) or propyloxycarbonyloxymethyl (POC), the
PSMA inhibitor is formulated for oral administration. In a
particular embodiment, the PSMA inhibitor formulated for oral
administration for use within the methods of the presently
disclosed subject matter is Tris-POC-2-PMPA. In a particular
embodiment, the PSMA inhibitor is formulated for administration via
a suppository. In a particular embodiment, the PSMA inhibitor
formulated for administration via a suppository is Tris-POC-2-PMPA.
In some embodiments, the suppository is a rectal suppository. The
agents may be delivered, for example, in a timed- or sustained-low
release form as is known to those skilled in the art. Techniques
for formulation and administration may be found in Remington: The
Science and Practice of Pharmacy (20.sup.th ed.) Lippincott,
Williams & Wilkins (2000).
[0049] Pharmaceutical preparations for oral use can be obtained by
combining the active compounds with solid excipients, optionally
grinding a resulting mixture, and processing the mixture of
granules, after adding suitable auxiliaries, if desired, to obtain
tablets or dragee cores. Suitable excipients are, in particular,
fillers such as sugars, including lactose, sucrose, mannitol, or
sorbitol; cellulose preparations, for example, maize starch, wheat
starch, rice starch, potato starch, gelatin, gum tragacanth, methyl
cellulose, hydroxypropylmethyl-cellulose, sodium
carboxymethyl-cellulose (CMC), and/or polyvinylpyrrolidone (PVP:
povidone). If desired, disintegrating agents may be added, such as
the cross-linked polyvinylpyrrolidone, agar, or alginic acid or a
salt thereof such as sodium alginate.
[0050] Dragee cores are provided with suitable coatings. For this
purpose, concentrated sugar solutions may be used, which may
optionally contain gum arabic, talc, polyvinylpyrrolidone, carbopol
gel, polyethylene glycol (PEG), and/or titanium dioxide, lacquer
solutions, and suitable organic solvents or solvent mixtures.
Dye-stuffs or pigments may be added to the tablets or dragee
coatings for identification or to characterize different
combinations of active compound doses.
[0051] Pharmaceutical preparations that can be used orally include
push-fit capsules made of gelatin, as well as soft, sealed capsules
made of gelatin, and a plasticizer, such as glycerol or sorbitol.
The push-fit capsules can contain the active ingredients in
admixture with filler such as lactose, binders such as starches,
and/or lubricants such as talc or magnesium stearate and,
optionally, stabilizers. In soft capsules, the active compounds may
be dissolved or suspended in suitable liquids, such as fatty oils,
liquid paraffin, or liquid polyethylene glycols (PEGs). In
addition, stabilizers may be added.
[0052] In another embodiment, where the PSMA inhibitor for use
within the methods of the presently disclosed subject matter is
2-PMPA or E2072, the PSMA inhibitor may be formulated into liquid
or solid dosage forms and administered systemically or locally.
Suitable routes may include rectal (e.g., via a suppository),
intestinal, or intraperitoneal delivery. Other suitable routes may
include various forms of parenteral delivery, including
intramuscular, subcutaneous, intramedullary injections, as well as
intrathecal, direct intraventricular, intravenous,
intra-articullar, intra-sternal, intra-synovial, intra-hepatic,
intralesional, intracranial, intraperitoneal, intranasal, or
intraocular injections or other modes of delivery.
[0053] For injection, the agents of the disclosure may be
formulated and diluted in aqueous solutions, such as in
physiologically compatible buffers such as Hank's solution,
Ringer's solution, or physiological saline buffer. For such
transmucosal administration, penetrants appropriate to the barrier
to be permeated are used in the formulation. Such penetrants are
generally known in the art.
[0054] Use of pharmaceutically acceptable inert carriers to
formulate the compounds herein disclosed for the practice of the
disclosure into dosages suitable for systemic administration is
within the scope of the disclosure. With proper choice of carrier
and suitable manufacturing practice, the compositions of the
present disclosure, in particular, those formulated as solutions,
may be administered parenterally, such as by intravenous injection.
The compounds can be formulated readily using pharmaceutically
acceptable carriers well known in the art into dosages suitable for
oral administration. Such carriers enable the compounds of the
disclosure to be formulated as tablets, pills, capsules, liquids,
gels, syrups, slurries, suspensions and the like, for oral
ingestion by a subject (e.g., patient) to be treated.
[0055] The compounds according to the disclosure are effective over
a wide dosage range. For example, in the treatment of adult humans,
dosages from 0.01 to 1000 mg, from 0.5 to 100 mg, from 1 to 50 mg
per day, and from 5 to 40 mg per day are examples of dosages that
may be used. A non-limiting dosage is 10 to 30 mg per day. The
exact dosage will depend upon the route of administration, the form
in which the compound is administered, the subject to be treated,
the body weight of the subject to be treated, and the preference
and experience of the attending physician.
[0056] Pharmaceutically acceptable salts are generally well known
to those of ordinary skill in the art, and may include, by way of
example but not limitation, acetate, benzenesulfonate, besylate,
benzoate, bicarbonate, bitartrate, bromide, calcium edetate,
carnsylate, carbonate, citrate, edetate, edisylate, estolate,
esylate, fumarate, gluceptate, gluconate, glutamate,
glycollylarsanilate, hexylresorcinate, hydrabamine, hydrobromide,
hydrochloride, hydroxynaphthoate, iodide, isethionate, lactate,
lactobionate, malate, maleate, mandelate, mesylate, mucate,
napsylate, nitrate, pamoate (embonate), pantothenate,
phosphate/diphosphate, polygalacturonate, salicylate, stearate,
subacetate, succinate, sulfate, tannate, tartrate, or teoclate.
Other pharmaceutically acceptable salts may be found in, for
example, Remington: The Science and Practice of Pharmacy (20.sup.th
ed.) Lippincott, Williams & Wilkins (2000). Pharmaceutically
acceptable salts include, for example, acetate, benzoate, bromide,
carbonate, citrate, gluconate, hydrobromide, hydrochloride,
maleate, mesylate, napsylate, pamoate (embonate), phosphate,
salicylate, succinate, sulfate, or tartrate.
[0057] Pharmaceutical compositions suitable for use in the present
disclosure include compositions wherein the active ingredients are
contained in an effective amount to achieve its intended purpose.
Determination of the effective amounts is well within the
capability of those skilled in the art, especially in light of the
detailed disclosure provided herein.
[0058] In addition to the active ingredients, these pharmaceutical
compositions may contain suitable pharmaceutically acceptable
carriers comprising excipients and auxiliaries which facilitate
processing of the active compounds into preparations which can be
used pharmaceutically. The preparations formulated for oral
administration may be in the form of tablets, dragees, capsules, or
solutions.
[0059] Additional therapeutic agents, which are normally
administered to treat or prevent IBD, may be administered together
with the PSMA inhibitors within the methods of the presently
disclosed subject matter. For example, anti-tumor necrosis factor
(TNF)-based therapies, such as infliximab (IFX), adalimumab and
certolizumab pegol, may be combined with the PSMA inhibitors of
this disclosure to treat IBD.
[0060] Other examples of agents with which the disclosed PSMA
inhibitors may also be combined include, without limitation,
anti-inflammatory agents such as corticosteroids, TNF blockers,
IL-I RA, azathioprine, cyclophosphamide, and sulfasalazine;
immunomodulatory and immunosuppressive agents such as cyclosporin,
tacrolimus, rapamycin, mycophenolate mofetil, interferons,
corticosteroids, cyclophophamide, azathioprine, and sulfasalazine;
neurotrophic factors, such as acetylcholinesterase inhibitors, MAO
inhibitors, interferons, anti-convulsants, ion channel blockers,
riluzole, and antiparkinsonian agents; agents for treating
cardiovascular disease such as beta-blockers, ACE inhibitors,
diuretics, nitrates, calcium channel blockers, and statins; agents
for treating liver disease such as corticosteroids, cholestyramine,
interferons, and anti-viral agents; agents for treating blood
disorders, such as corticosteroids, anti-leukemic agents, and
growth factors; agents for treating diabetes such as insulin,
insulin analogues, alpha glucosidase inhibitors, biguanides, and
insulin sensitizers; and agents for treating immunodeficiency
disorders such as gamma globulin.
[0061] These additional agents may be administered separately, as
part of a multiple dosage regimen, from the inhibitor-containing
composition. Alternatively, these agents may be part of a single
dosage form, mixed together with the inhibitor in a single
composition.
III. Definitions
[0062] Although specific terms are employed herein, they are used
in a generic and descriptive sense only and not for purposes of
limitation. Unless otherwise defined, all technical and scientific
terms used herein have the same meaning as commonly understood by
one of ordinary skill in the art to which this presently described
subject matter belongs.
[0063] While the following terms in relation to PSMA inhibitors are
believed to be well understood by one of ordinary skill in the art,
the following definitions are set forth to facilitate explanation
of the presently disclosed subject matter. These definitions are
intended to supplement and illustrate, not preclude, the
definitions that would be apparent to one of ordinary skill in the
art upon review of the present disclosure.
[0064] The terms substituted, whether preceded by the term
"optionally" or not, and substituent, as used herein, refer to the
ability, as appreciated by one skilled in this art, to change one
functional group for another functional group provided that the
valency of all atoms is maintained. When more than one position in
any given structure may be substituted with more than one
substituent selected from a specified group, the substituent may be
either the same or different at every position. The substituents
also may be further substituted (e.g., an aryl group substituent
may have another substituent off it, such as another aryl group,
which is further substituted, for example, with fluorine at one or
more positions).
[0065] Where substituent groups or linking groups are specified by
their conventional chemical formulae, written from left to right,
they equally encompass the chemically identical substituents that
would result from writing the structure from right to left, e.g.,
--CH.sub.2O-- is equivalent to --OCH.sub.2--; --C(.dbd.O)O-- is
equivalent to --OC(.dbd.O)--; --OC(.dbd.O)NR-- is equivalent to
--NRC(.dbd.O)O--, and the like.
[0066] When the term "independently selected" is used, the
substituents being referred to (e.g., R groups, such as groups
R.sub.1, R.sub.2, and the like, or variables, such as "m" and "n"),
can be identical or different. For example, both R.sub.1 and
R.sub.2 can be substituted alkyls, or R.sub.1 can be hydrogen and
R.sub.2 can be a substituted alkyl, and the like.
[0067] The terms "a," "an," or "a(n)," when used in reference to a
group of substituents herein, mean at least one. For example, where
a compound is substituted with "an" alkyl or aryl, the compound is
optionally substituted with at least one alkyl and/or at least one
aryl. Moreover, where a moiety is substituted with an R
substituent, the group may be referred to as "R-substituted." Where
a moiety is R-substituted, the moiety is substituted with at least
one R substituent and each R substituent is optionally
different.
[0068] A named "R" or group will generally have the structure that
is recognized in the art as corresponding to a group having that
name, unless specified otherwise herein. For the purposes of
illustration, certain representative "R" groups as set forth above
are defined below.
[0069] Description of compounds of the present disclosure is
limited by principles of chemical bonding known to those skilled in
the art. Accordingly, where a group may be substituted by one or
more of a number of substituents, such substitutions are selected
so as to comply with principles of chemical bonding and to give
compounds which are not inherently unstable and/or would be known
to one of ordinary skill in the art as likely to be unstable under
ambient conditions, such as aqueous, neutral, and several known
physiological conditions. For example, a heterocycloalkyl or
heteroaryl is attached to the remainder of the molecule via a ring
heteroatom in compliance with principles of chemical bonding known
to those skilled in the art thereby avoiding inherently unstable
compounds.
[0070] The term hydrocarbon, as used herein, refers to any chemical
group comprising hydrogen and carbon. The hydrocarbon may be
substituted or unsubstituted. As would be known to one skilled in
this art, all valencies must be satisfied in making any
substitutions. The hydrocarbon may be unsaturated, saturated,
branched, unbranched, cyclic, polycyclic, or heterocyclic.
Illustrative hydrocarbons are further defined herein below and
include, for example, methyl, ethyl, n-propyl, iso-propyl,
cyclopropyl, allyl, vinyl, n-butyl, tert-butyl, ethynyl,
cyclohexyl, methoxy, diethylamino, and the like.
[0071] The term "alkyl," by itself or as part of another
substituent, means, unless otherwise stated, a straight (i.e.,
unbranched) or branched chain, acyclic or cyclic hydrocarbon group,
or combination thereof, which may be fully saturated, mono- or
polyunsaturated and can include di- and multivalent groups, having
the number of carbon atoms designated (i.e., C.sub.1-C.sub.10 means
one to ten carbons). In particular embodiments, the term "alkyl"
refers to C.sub.1-20 inclusive, linear (i.e., "straight-chain"),
branched, or cyclic, saturated or at least partially and in some
cases fully unsaturated (i.e., alkenyl and alkynyl) hydrocarbon
radicals derived from a hydrocarbon moiety containing between one
and twenty carbon atoms by removal of a single hydrogen atom.
[0072] Representative saturated hydrocarbon groups include, but are
not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl,
isobutyl, sec-butyl, tert-butyl, n-pentyl, sec-pentyl, iso-pentyl,
neopentyl, n-hexyl, sec-hexyl, n-heptyl, n-octyl, n-decyl,
n-undecyl, dodecyl, cyclohexyl, (cyclohexyl)methyl,
cyclopropylmethyl, and homologs and isomers thereof "Branched"
refers to an alkyl group in which a lower alkyl group, such as
methyl, ethyl or propyl, is attached to a linear alkyl chain.
"Lower alkyl" refers to an alkyl group having 1 to about 8 carbon
atoms (i.e., a C.sub.1-8 alkyl), e.g., 1, 2, 3, 4, 5, 6, 7, or 8
carbon atoms. "Higher alkyl" refers to an alkyl group having about
10 to about 20 carbon atoms, e.g., 10, 11, 12, 13, 14, 15, 16, 17,
18, 19, or 20 carbon atoms. In certain embodiments, "alkyl" refers,
in particular, to C.sub.1-8 straight-chain alkyls. In other
embodiments, "alkyl" refers, in particular, to C.sub.1-8
branched-chain alkyls.
[0073] Alkyl groups can optionally be substituted (a "substituted
alkyl") with one or more alkyl group substituents, which can be the
same or different. The term "alkyl group substituent" includes but
is not limited to alkyl, substituted alkyl, halo, arylamino, acyl,
hydroxyl, aryloxyl, alkoxyl, alkylthio, arylthio, aralkyloxyl,
aralkylthio, carboxyl, alkoxycarbonyl, oxo, and cycloalkyl. There
can be optionally inserted along the alkyl chain one or more
oxygen, sulfur or substituted or unsubstituted nitrogen atoms,
wherein the nitrogen substituent is hydrogen, lower alkyl (also
referred to herein as "alkylaminoalkyl"), or aryl.
[0074] Thus, as used herein, the term "substituted alkyl" includes
alkyl groups, as defined herein, in which one or more atoms or
functional groups of the alkyl group are replaced with another atom
or functional group, including for example, alkyl, substituted
alkyl, halogen, aryl, substituted aryl, alkoxyl, hydroxyl, nitro,
amino, alkylamino, dialkylamino, sulfate, and mercapto.
[0075] The term "heteroalkyl," by itself or in combination with
another term, means, unless otherwise stated, a stable straight or
branched chain, or cyclic hydrocarbon group, or combinations
thereof, consisting of at least one carbon atoms and at least one
heteroatom selected from the group consisting of O, N, P, Si and S,
and wherein the nitrogen, phosphorus, and sulfur atoms may
optionally be oxidized and the nitrogen heteroatom may optionally
be quaternized. The heteroatom(s) O, N, P and S and Si may be
placed at any interior position of the heteroalkyl group or at the
position at which alkyl group is attached to the remainder of the
molecule. Examples include, but are not limited to,
--CH.sub.2--CH.sub.2--O--CH.sub.3,
--CH.sub.2--CH.sub.2--NH--CH.sub.3,
--CH.sub.2--CH.sub.2--N(CH.sub.3)--CH.sub.3,
--CH.sub.2--S--CH.sub.2--CH.sub.3,
--CH.sub.2--CH.sub.25--S(O)--CH.sub.3,
--CH.sub.2--CH.sub.2--S(O).sub.2--CH.sub.3,
--CH.dbd.CH--O--CH.sub.3, --Si(CH.sub.3).sub.3,
--CH.sub.2--CH.dbd.N--OCH.sub.3,
--CH.dbd.CH--N(CH.sub.3)--CH.sub.3, O--CH.sub.3,
--O--CH.sub.2--CH.sub.3, and --CN. Up to two or three heteroatoms
may be consecutive, such as, for example, --CH.sub.2--NH--OCH.sub.3
and --CH.sub.2--O--Si(CH.sub.3).sub.3.
[0076] As described above, heteroalkyl groups, as used herein,
include those groups that are attached to the remainder of the
molecule through a heteroatom, such as --C(O)R', --C(O)NR',
--NR'R'', --OR', --SR, and/or --SO.sub.2R'. Where "heteroalkyl" is
recited, followed by recitations of specific heteroalkyl groups,
such as --NR'R or the like, it will be understood that the terms
heteroalkyl and --NR'R'' are not redundant or mutually exclusive.
Rather, the specific heteroalkyl groups are recited to add clarity.
Thus, the term "heteroalkyl" should not be interpreted herein as
excluding specific heteroalkyl groups, such as --NR'R'' or the
like.
[0077] "Cyclic" and "cycloalkyl" refer to a non-aromatic mono- or
multicyclic ring system of about 3 to about 10 carbon atoms, e.g.,
3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. The cycloalkyl group can
be optionally partially unsaturated. The cycloalkyl group also can
be optionally substituted with an alkyl group substituent as
defined herein, oxo, and/or alkylene. There can be optionally
inserted along the cyclic alkyl chain one or more oxygen, sulfur or
substituted or unsubstituted nitrogen atoms, wherein the nitrogen
substituent is hydrogen, alkyl, substituted alkyl, aryl, or
substituted aryl, thus providing a heterocyclic group.
Representative monocyclic cycloalkyl rings include cyclopentyl,
cyclohexyl, and cycloheptyl. Multicyclic cycloalkyl rings include
adamantyl, octahydronaphthyl, decalin, camphor, camphane, and
noradamantyl, and fused ring systems, such as dihydro- and
tetrahydronaphthalene, and the like.
[0078] The term "cycloalkylalkyl," as used herein, refers to a
cycloalkyl group as defined hereinabove, which is attached to the
parent molecular moiety through an alkyl group, also as defined
above. Examples of cycloalkylalkyl groups include cyclopropylmethyl
and cyclopentylethyl.
[0079] The terms "cycloheteroalkyl" or "heterocycloalkyl" refer to
a non-aromatic ring system, unsaturated or partially unsaturated
ring system, such as a 3- to 10-member substituted or unsubstituted
cycloalkyl ring system, including one or more heteroatoms, which
can be the same or different, and are selected from the group
consisting of nitrogen (N), oxygen (O), sulfur (S), phosphorus (P),
and silicon (Si), and optionally can include one or more double
bonds.
[0080] The cycloheteroalkyl ring can be optionally fused to or
otherwise attached to other cycloheteroalkyl rings and/or
non-aromatic hydrocarbon rings. Heterocyclic rings include those
having from one to three heteroatoms independently selected from
oxygen, sulfur, and nitrogen, in which the nitrogen and sulfur
heteroatoms may optionally be oxidized and the nitrogen heteroatom
may optionally be quaternized. In certain embodiments, the term
heterocylic refers to a non-aromatic 5-, 6-, or 7-membered ring or
a polycyclic group wherein at least one ring atom is a heteroatom
selected from O, S, and N (wherein the nitrogen and sulfur
heteroatoms may be optionally oxidized), including, but not limited
to, a bi- or tri-cyclic group, comprising fused six-membered rings
having between one and three heteroatoms independently selected
from the oxygen, sulfur, and nitrogen, wherein (i) each 5-membered
ring has 0 to 2 double bonds, each 6-membered ring has 0 to 2
double bonds, and each 7-membered ring has 0 to 3 double bonds,
(ii) the nitrogen and sulfur heteroatoms may be optionally
oxidized, (iii) the nitrogen heteroatom may optionally be
quaternized, and (iv) any of the above heterocyclic rings may be
fused to an aryl or heteroaryl ring. Representative
cycloheteroalkyl ring systems include, but are not limited to
pyrrolidinyl, pyrrolinyl, imidazolidinyl, imidazolinyl,
pyrazolidinyl, pyrazolinyl, piperidyl, piperazinyl, indolinyl,
quinuclidinyl, morpholinyl, thiomorpholinyl, thiadiazinanyl,
tetrahydrofuranyl, and the like.
[0081] The terms "cycloalkyl" and "heterocycloalkyl", by themselves
or in combination with other terms, represent, unless otherwise
stated, cyclic versions of "alkyl" and "heteroalkyl", respectively.
Additionally, for heterocycloalkyl, a heteroatom can occupy the
position at which the heterocycle is attached to the remainder of
the molecule. Examples of cycloalkyl include, but are not limited
to, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl,
cycloheptyl, and the like. Examples of heterocycloalkyl include,
but are not limited to, 1-(1,2,5,6-tetrahydropyridyl),
1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl,
3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl,
tetrahydrothien-2-yl, tetrahydrothien-3-yl, 1-piperazinyl,
2-piperazinyl, and the like. The terms "cycloalkylene" and
"heterocycloalkylene" refer to the divalent derivatives of
cycloalkyl and heterocycloalkyl, respectively.
[0082] An unsaturated alkyl group is one having one or more double
bonds or triple bonds. Examples of unsaturated alkyl groups
include, but are not limited to, vinyl, 2-propenyl, crotyl,
2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl,
3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and the
higher homologs and isomers. Alkyl groups which are limited to
hydrocarbon groups are termed "homoalkyl."
[0083] More particularly, the term "alkenyl" as used herein refers
to a monovalent group derived from a C.sub.1-20 inclusive straight
or branched hydrocarbon moiety having at least one carbon-carbon
double bond by the removal of a single hydrogen atom. Alkenyl
groups include, for example, ethenyl (i.e., vinyl), propenyl,
butenyl, 1-methyl-2-buten-1-yl, pentenyl, hexenyl, octenyl, and
butadienyl.
[0084] The term "cycloalkenyl" as used herein refers to a cyclic
hydrocarbon containing at least one carbon-carbon double bond.
Examples of cycloalkenyl groups include cyclopropenyl,
cyclobutenyl, cyclopentenyl, cyclopentadiene, cyclohexenyl,
1,3-cyclohexadiene, cycloheptenyl, cycloheptatrienyl, and
cyclooctenyl.
[0085] The term "alkynyl" as used herein refers to a monovalent
group derived from a straight or branched C.sub.1-20 hydrocarbon of
a designed number of carbon atoms containing at least one
carbon-carbon triple bond. Examples of "alkynyl" include ethynyl,
2-propynyl (propargyl), 1-propynyl, pentynyl, hexynyl, heptynyl,
and allenyl groups, and the like.
[0086] The term "alkylene" by itself or a part of another
substituent refers to a straight or branched bivalent aliphatic
hydrocarbon group derived from an alkyl group having from 1 to
about 20 carbon atoms, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,
13, 14, 15, 16, 17, 18, 19, or 20 carbon atoms. The alkylene group
can be straight, branched or cyclic. The alkylene group also can be
optionally unsaturated and/or substituted with one or more "alkyl
group substituents." There can be optionally inserted along the
alkylene group one or more oxygen, sulfur or substituted or
unsubstituted nitrogen atoms (also referred to herein as
"alkylaminoalkyl"), wherein the nitrogen substituent is alkyl as
previously described. Exemplary alkylene groups include methylene
(--CH.sub.2--); ethylene (--CH.sub.2--CH.sub.2--); propylene
(--(CH.sub.2).sub.3--); cyclohexylene (--C.sub.6H.sub.10--);
--CH.dbd.CH--CH.dbd.CH--; --CH.dbd.CH--CH.sub.2--;
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2--,
--CH.sub.2CH.dbd.CHCH.sub.2--, --CH.sub.2CsCCH.sub.2--,
--CH.sub.2CH.sub.2CH(CH.sub.2CH.sub.2CH.sub.3)CH.sub.2--,
--(CH.sub.2).sub.q--N(R)--(CH.sub.2).sub.r--, wherein each of q and
r is independently an integer from 0 to about 20, e.g., 0, 1, 2, 3,
4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20,
and R is hydrogen or lower alkyl; methylenedioxyl
(--O--CH.sub.2--O--); and ethylenedioxyl
(--O--(CH.sub.2).sub.2--O--). An alkylene group can have about 2 to
about 3 carbon atoms and can further have 6-20 carbons. Typically,
an alkyl (or alkylene) group will have from 1 to 24 carbon atoms,
with those groups having 10 or fewer carbon atoms being some
embodiments of the present disclosure. A "lower alkyl" or "lower
alkylene" is a shorter chain alkyl or alkylene group, generally
having eight or fewer carbon atoms.
[0087] The term "heteroalkylene" by itself or as part of another
substituent means a divalent group derived from heteroalkyl, as
exemplified, but not limited by,
--CH.sub.2--CH.sub.2--S--CH.sub.2--CH.sub.2-- and
--CH.sub.2--S--CH.sub.2--CH.sub.2--NH--CH.sub.2--. For
heteroalkylene groups, heteroatoms can also occupy either or both
of the chain termini (e.g., alkyleneoxo, alkylenedioxo,
alkyleneamino, alkylenediamino, and the like). Still further, for
alkylene and heteroalkylene linking groups, no orientation of the
linking group is implied by the direction in which the formula of
the linking group is written. For example, the formula --C(O)OR'--
represents both --C(O)OR'-- and --R'OC(O)--.
[0088] The term "aryl" means, unless otherwise stated, an aromatic
hydrocarbon substituent that can be a single ring or multiple rings
(such as from 1 to 3 rings), which are fused together or linked
covalently. The term "heteroaryl" refers to aryl groups (or rings)
that contain from one to four heteroatoms (in each separate ring in
the case of multiple rings) selected from N, O, and S, wherein the
nitrogen and sulfur atoms are optionally oxidized, and the nitrogen
atom(s) are optionally quaternized. A heteroaryl group can be
attached to the remainder of the molecule through a carbon or
heteroatom. Non-limiting examples of aryl and heteroaryl groups
include phenyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl,
2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl,
pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl,
3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl,
5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl,
3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl,
purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl,
2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, and 6-quinolyl.
Substituents for each of above noted aryl and heteroaryl ring
systems are selected from the group of acceptable substituents
described below. The terms "arylene" and "heteroarylene" refer to
the divalent forms of aryl and heteroaryl, respectively.
[0089] For brevity, the term "aryl" when used in combination with
other terms (e.g., aryloxo, arylthioxo, arylalkyl) includes both
aryl and heteroaryl rings as defined above. Thus, the terms
"arylalkyl" and "heteroarylalkyl" are meant to include those groups
in which an aryl or heteroaryl group is attached to an alkyl group
(e.g., benzyl, phenethyl, pyridylmethyl, furylmethyl, and the like)
including those alkyl groups in which a carbon atom (e.g., a
methylene group) has been replaced by, for example, an oxygen atom
(e.g., phenoxymethyl, 2-pyridyloxymethyl, 3-(1-naphthyloxy)propyl,
and the like). However, the term "haloaryl," as used herein is
meant to cover only aryls substituted with one or more
halogens.
[0090] Where a heteroalkyl, heterocycloalkyl, or heteroaryl
includes a specific number of members (e.g. "3 to 7 membered"), the
term "member" refers to a carbon or heteroatom.
[0091] Further, a structure represented generally by the
formula:
##STR00007##
as used herein refers to a ring structure, for example, but not
limited to a 3-carbon, a 4-carbon, a 5-carbon, a 6-carbon, a
7-carbon, and the like, aliphatic and/or aromatic cyclic compound,
including a saturated ring structure, a partially saturated ring
structure, and an unsaturated ring structure, comprising a
substituent R group, wherein the R group can be present or absent,
and when present, one or more R groups can each be substituted on
one or more available carbon atoms of the ring structure. The
presence or absence of the R group and number of R groups is
determined by the value of the variable "n," which is an integer
generally having a value ranging from 0 to the number of carbon
atoms on the ring available for substitution. Each R group, if more
than one, is substituted on an available carbon of the ring
structure rather than on another R group. For example, the
structure above where n is 0 to 2 would comprise compound groups
including, but not limited to:
##STR00008##
and the like.
[0092] A dashed line representing a bond in a cyclic ring structure
indicates that the bond can be either present or absent in the
ring. That is, a dashed line representing a bond in a cyclic ring
structure indicates that the ring structure is selected from the
group consisting of a saturated ring structure, a partially
saturated ring structure, and an unsaturated ring structure.
[0093] The symbol () denotes the point of attachment of a moiety to
the remainder of the molecule.
[0094] When a named atom of an aromatic ring or a heterocyclic
aromatic ring is defined as being "absent," the named atom is
replaced by a direct bond.
[0095] Each of above terms (e.g., "alkyl," "heteroalkyl,"
"cycloalkyl, and "heterocycloalkyl", "aryl," "heteroaryl,"
"phosphonate," and "sulfonate" as well as their divalent
derivatives) are meant to include both substituted and
unsubstituted forms of the indicated group. Optional substituents
for each type of group are provided below.
[0096] Substituents for alkyl, heteroalkyl, cycloalkyl,
heterocycloalkyl monovalent and divalent derivative groups
(including those groups often referred to as alkylene, alkenyl,
heteroalkylene, heteroalkenyl, alkynyl, cycloalkyl,
heterocycloalkyl, cycloalkenyl, and heterocycloalkenyl) can be one
or more of a variety of groups selected from, but not limited to:
--OR', .dbd.O, .dbd.NR', .dbd.N--OR', --NR'R'', --SR', -halogen,
--SiR'R''R''', --OC(O)R', --C(O)R', --CO.sub.2R', --C(O)NR'R'',
--OC(O)NR'R'', --NR''C(O)R', --NR'--C(O)NR''R''', --NR''C(O)OR',
--NR--C(NR'R'').dbd.NR''', --S(O)R', --S(O).sub.2R',
--S(O).sub.2NR'R'', --NRSO.sub.2R', --CN and --NO.sub.2 in a number
ranging from zero to (2m'+1), where m' is the total number of
carbon atoms in such groups. R', R'', R''' and R'''' each may
independently refer to hydrogen, substituted or unsubstituted
heteroalkyl, substituted or unsubstituted cycloalkyl, substituted
or unsubstituted heterocycloalkyl, substituted or unsubstituted
aryl (e.g., aryl substituted with 1-3 halogens), substituted or
unsubstituted alkyl, alkoxy or thioalkoxy groups, or arylalkyl
groups. As used herein, an "alkoxy" group is an alkyl attached to
the remainder of the molecule through a divalent oxygen. When a
compound of the disclosure includes more than one R group, for
example, each of the R groups is independently selected as are each
R', R'', R''' and R'''' groups when more than one of these groups
is present. When R' and R'' are attached to the same nitrogen atom,
they can be combined with the nitrogen atom to form a 4-, 5-, 6-,
or 7-membered ring. For example, --NR'R'' is meant to include, but
not be limited to, 1-pyrrolidinyl and 4-morpholinyl. From the above
discussion of substituents, one of skill in the art will understand
that the term "alkyl" is meant to include groups including carbon
atoms bound to groups other than hydrogen groups, such as haloalkyl
(e.g., --CF.sub.3 and --CH.sub.2CF.sub.3) and acyl (e.g.,
--C(O)CH.sub.3, --C(O)CF.sub.3, --C(O)CH.sub.2OCH.sub.3, and the
like).
[0097] Similar to the substituents described for alkyl groups
above, exemplary substituents for aryl and heteroaryl groups (as
well as their divalent derivatives) are varied and are selected
from, for example: halogen, --OR', --NR'R'', --SR', -halogen,
--SiR'R''R'', --OC(O)R', --C(O)R', --CO.sub.2R', --C(O)NR'R'',
--OC(O)NR'R'', --NR''C(O)R', --NR'--C(O)NR''R''', --NR''C(O)OR',
--NR--C(NR'R''R''').dbd.NR'''', --NR--C(NR'R'').dbd.NR'''--S(O)R',
--S(O).sub.2R', --S(O).sub.2NR'R'', --NRSO.sub.2R', --CN and
--NO.sub.2, --R', --N.sub.3, --CH(Ph).sub.2,
fluoro(C.sub.1-C.sub.4)alkoxo, and fluoro(C.sub.1-C.sub.4)alkyl, in
a number ranging from zero to the total number of open valences on
aromatic ring system; and where R', R'', R''' and R'''' may be
independently selected from hydrogen, substituted or unsubstituted
alkyl, substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl and substituted
or unsubstituted heteroaryl. When a compound of the disclosure
includes more than one R group, for example, each of the R groups
is independently selected as are each R', R'', R''' and R''''
groups when more than one of these groups is present.
[0098] Two of the substituents on adjacent atoms of aryl or
heteroaryl ring may optionally form a ring of the formula
-T-C(O)--(CRR').sub.q--U--, wherein T and U are independently
--NR--, --O--, --CRR'-- or a single bond, and q is an integer of
from 0 to 3. Alternatively, two of the substituents on adjacent
atoms of aryl or heteroaryl ring may optionally be replaced with a
substituent of the formula -A-(CH.sub.2).sub.r--B--, wherein A and
B are independently --CRR'--, --O--, --NR--, --S--, --S(O)--,
--S(O).sub.2--, --S(O).sub.2NR'-- or a single bond, and r is an
integer of from 1 to 4.
[0099] One of the single bonds of the new ring so formed may
optionally be replaced with a double bond. Alternatively, two of
the substituents on adjacent atoms of aryl or heteroaryl ring may
optionally be replaced with a substituent of the formula
--(CRR').sub.s--X'--(C''R''').sub.d--, where s and d are
independently integers of from 0 to 3, and X' is --O--, --NR'--,
--S--, --S(O)--, --S(O).sub.2--, or --S(O).sub.2NR'--. The
substituents R, R', R'' and R''' may be independently selected from
hydrogen, substituted or unsubstituted alkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, and
substituted or unsubstituted heteroaryl.
[0100] As used herein, the term "acyl" refers to an organic acid
group wherein the --OH of the carboxyl group has been replaced with
another substituent and has the general formula RC(.dbd.O)--,
wherein R is an alkyl, alkenyl, alkynyl, aryl, carbocylic,
heterocyclic, or aromatic heterocyclic group as defined herein). As
such, the term "acyl" specifically includes arylacyl groups, such
as an acetylfuran and a phenacyl group. Specific examples of acyl
groups include acetyl and benzoyl.
[0101] The terms "alkoxyl" or "alkoxy" are used interchangeably
herein and refer to a saturated (i.e., alkyl-O--) or unsaturated
(i.e., alkenyl-O-- and alkynyl-O--) group attached to the parent
molecular moiety through an oxygen atom, wherein the terms "alkyl,"
"alkenyl," and "alkynyl" are as previously described and can
include C.sub.1-20 inclusive, linear, branched, or cyclic,
saturated or unsaturated oxo-hydrocarbon chains, including, for
example, methoxyl, ethoxyl, propoxyl, isopropoxyl, n-butoxyl,
sec-butoxyl, t-butoxyl, and n-pentoxyl, neopentoxy, n-hexoxy, and
the like.
[0102] The term "alkoxyalkyl" as used herein refers to an
alkyl-O-alkyl ether, for example, a methoxyethyl or an ethoxymethyl
group.
[0103] "Aryloxyl" refers to an aryl-O-- group wherein the aryl
group is as previously described, including a substituted aryl. The
term "aryloxyl" as used herein can refer to phenyloxyl or
hexyloxyl, and alkyl, substituted alkyl, halo, or alkoxyl
substituted phenyloxyl or hexyloxyl.
[0104] "Aralkyl" refers to an aryl-alkyl-group wherein aryl and
alkyl are as previously described, and included substituted aryl
and substituted alkyl. Exemplary aralkyl groups include benzyl,
phenylethyl, and naphthylmethyl.
[0105] "Aralkyloxyl" refers to an aralkyl-O-- group wherein the
aralkyl group is as previously described. An exemplary aralkyloxyl
group is benzyloxyl.
[0106] "Alkoxycarbonyl" refers to an alkyl-O--CO-- group. Exemplary
alkoxycarbonyl groups include methoxycarbonyl, ethoxycarbonyl,
butyloxycarbonyl, and t-butyloxycarbonyl.
[0107] "Aryloxycarbonyl" refers to an aryl-O--CO-- group. Exemplary
aryloxycarbonyl groups include phenoxy- and naphthoxy-carbonyl.
[0108] "Aralkoxycarbonyl" refers to an aralkyl-O--CO-- group. An
exemplary aralkoxycarbonyl group is benzyloxycarbonyl.
[0109] "Carbamoyl" refers to an amide group of the formula
--CONH.sub.2. "Alkylcarbamoyl" refers to a R'RN--CO-- group wherein
one of R and R' is hydrogen and the other of R and R' is alkyl
and/or substituted alkyl as previously described.
"Dialkylcarbamoyl" refers to a R'RN--CO-- group wherein each of R
and R' is independently alkyl and/or substituted alkyl as
previously described.
[0110] The term carbonyldioxyl, as used herein, refers to a
carbonate group of the formula --O--CO--OR.
[0111] "Acyloxyl" refers to an acyl-O-- group wherein acyl is as
previously described.
[0112] The term "amino" refers to the --NH.sub.2 group and also
refers to a nitrogen containing group as is known in the art
derived from ammonia by the replacement of one or more hydrogen
radicals by organic radicals. For example, the terms "acylamino"
and "alkylamino" refer to specific N-substituted organic radicals
with acyl and alkyl substituent groups respectively.
[0113] An "aminoalkyl" as used herein refers to an amino group
covalently bound to an alkylene linker. More particularly, the
terms alkylamino, dialkylamino, and trialkylamino as used herein
refer to one, two, or three, respectively, alkyl groups, as
previously defined, attached to the parent molecular moiety through
a nitrogen atom. The term alkylamino refers to a group having the
structure --NHR' wherein R' is an alkyl group, as previously
defined; whereas the term dialkylamino refers to a group having the
structure --NR'R'', wherein R' and R'' are each independently
selected from the group consisting of alkyl groups. The term
trialkylamino refers to a group having the structure --NR'R''R'',
wherein R', R'', and R' are each independently selected from the
group consisting of alkyl groups. Additionally, R', R'', and/or R'
taken together may optionally be --(CH.sub.2).sub.k-- where k is an
integer from 2 to 6. Examples include, but are not limited to,
methylamino, dimethylamino, ethylamino, diethylamino,
diethylaminocarbonyl, methylethylamino, iso-propylamino,
piperidino, trimethylamino, and propylamino.
[0114] The amino group is --NR'R'', wherein R' and R'' are
typically selected from hydrogen, substituted or unsubstituted
alkyl, substituted or unsubstituted heteroalkyl, substituted or
unsubstituted cycloalkyl, substituted or unsubstituted
heterocycloalkyl, substituted or unsubstituted aryl, or substituted
or unsubstituted heteroaryl.
[0115] The terms alkylthioether and thioalkoxyl refer to a
saturated (i.e., alkyl-S--) or unsaturated (i.e., alkenyl-S-- and
alkynyl-S--) group attached to the parent molecular moiety through
a sulfur atom. Examples of thioalkoxyl moieties include, but are
not limited to, methylthio, ethylthio, propylthio, isopropylthio,
n-butylthio, and the like.
[0116] "Acylamino" refers to an acyl-NH-- group wherein acyl is as
previously described. "Aroylamino" refers to an aroyl-NH-- group
wherein aroyl is as previously described.
[0117] The term "carbonyl" refers to the --(C.dbd.O)-- group.
[0118] The term "carboxyl" refers to the --COOH group. Such groups
also are referred to herein as a "carboxylic acid" moiety.
[0119] The terms "halo," "halide," or "halogen" as used herein
refer to fluoro, chloro, bromo, and iodo groups. Additionally,
terms such as "haloalkyl," are meant to include monohaloalkyl and
polyhaloalkyl. For example, the term "halo(C.sub.1-C.sub.4)alkyl"
is mean to include, but not be limited to, trifluoromethyl,
2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the
like.
[0120] The term "hydroxyl" refers to the --OH group.
[0121] The term "hydroxyalkyl" refers to an alkyl group substituted
with an --OH group.
[0122] The term "mercapto" refers to the --SH group.
[0123] The term "oxo" as used herein means an oxygen atom that is
double bonded to a carbon atom or to another element.
[0124] The term "nitro" refers to the --NO.sub.2 group.
[0125] The term "thio" refers to a compound described previously
herein wherein a carbon or oxygen atom is replaced by a sulfur
atom.
[0126] The term "sulfate" refers to the --SO.sub.4 group.
[0127] The term thiohydroxyl or thiol, as used herein, refers to a
group of the formula --SH.
[0128] The term ureido refers to a urea group of the formula
--NH--CO--NH.sub.2.
[0129] Unless otherwise explicitly defined, a "substituent group,"
as used herein, includes a functional group selected from one or
more of the following moieties, which are defined herein:
[0130] (A) --OH, --NH.sub.2, --SH, --CN, --CF.sub.3, --NO.sub.2,
oxo, halogen, unsubstituted alkyl, unsubstituted heteroalkyl,
unsubstituted cycloalkyl, unsubstituted heterocycloalkyl,
unsubstituted aryl, unsubstituted heteroaryl, and
[0131] (B) alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,
and heteroaryl, substituted with at least one substituent selected
from:
[0132] (i) oxo, --OH, --NH.sub.2, --SH, --CN, --CF.sub.3,
--NO.sub.2, halogen, unsubstituted alkyl, unsubstituted
heteroalkyl, unsubstituted cycloalkyl, unsubstituted
heterocycloalkyl, unsubstituted aryl, unsubstituted heteroaryl,
and
[0133] (ii) alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,
and heteroaryl, substituted with at least one substituent selected
from:
[0134] (a) oxo, --OH, --NH.sub.2, --SH, --CN, --CF.sub.3,
--NO.sub.2, halogen, unsubstituted alkyl, unsubstituted
heteroalkyl, unsubstituted cycloalkyl, unsubstituted
heterocycloalkyl, unsubstituted aryl, unsubstituted heteroaryl,
and
[0135] (b) alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,
or heteroaryl, substituted with at least one substituent selected
from oxo, --OH, --NH.sub.2, --SH, --CN, --CF.sub.3, --NO.sub.2,
halogen, unsubstituted alkyl, unsubstituted heteroalkyl,
unsubstituted cycloalkyl, unsubstituted heterocycloalkyl,
unsubstituted aryl, and unsubstituted heteroaryl.
[0136] A "lower substituent" or "lower substituent group," as used
herein means a group selected from all of the substituents
described hereinabove for a "substituent group," wherein each
substituted or unsubstituted alkyl is a substituted or
unsubstituted C.sub.1-C.sub.8 alkyl, each substituted or
unsubstituted heteroalkyl is a substituted or unsubstituted 2 to 8
membered heteroalkyl, each substituted or unsubstituted cycloalkyl
is a substituted or unsubstituted C.sub.5-C.sub.7 cycloalkyl, and
each substituted or unsubstituted heterocycloalkyl is a substituted
or unsubstituted 5 to 7 membered heterocycloalkyl.
[0137] A "size-limited substituent" or "size-limited substituent
group," as used herein means a group selected from all of the
substituents described above for a "substituent group," wherein
each substituted or unsubstituted alkyl is a substituted or
unsubstituted C.sub.1-C.sub.20 alkyl, each substituted or
unsubstituted heteroalkyl is a substituted or unsubstituted 2 to 20
membered heteroalkyl, each substituted or unsubstituted cycloalkyl
is a substituted or unsubstituted C.sub.4-C.sub.8 cycloalkyl, and
each substituted or unsubstituted heterocycloalkyl is a substituted
or unsubstituted 4 to 8 membered heterocycloalkyl.
[0138] Throughout the specification and claims, a given chemical
formula or name shall encompass all tautomers, congeners, and
optical- and stereoisomers, as well as racemic mixtures where such
isomers and mixtures exist.
[0139] Certain compounds of the present disclosure possess
asymmetric carbon atoms (optical or chiral centers) or double
bonds; the enantiomers, racemates, diastereomers, tautomers,
geometric isomers, stereoisometric forms that may be defined, in
terms of absolute stereochemistry, as (R)- or (S)- or, as (D)- or
(L)- for amino acids, and individual isomers are encompassed within
the scope of the present disclosure. The compounds of the present
disclosure do not include those which are known in art to be too
unstable to synthesize and/or isolate. The present disclosure is
meant to include compounds in racemic and optically pure forms.
Optically active (R)- and (S)-, or (D)- and (L)-isomers may be
prepared using chiral synthons or chiral reagents, or resolved
using conventional techniques. When the compounds described herein
contain olefenic bonds or other centers of geometric asymmetry, and
unless specified otherwise, it is intended that the compounds
include both E and Z geometric isomers.
[0140] Unless otherwise stated, structures depicted herein are also
meant to include all stereochemical forms of the structure; i.e.,
the R and S configurations for each asymmetric center. Therefore,
single stereochemical isomers as well as enantiomeric and
diastereomeric mixtures of the present compounds are within the
scope of the disclosure.
[0141] It will be apparent to one skilled in the art that certain
compounds of this disclosure may exist in tautomeric forms, all
such tautomeric forms of the compounds being within the scope of
the disclosure. The term "tautomer," as used herein, refers to one
of two or more structural isomers which exist in equilibrium and
which are readily converted from one isomeric form to another.
[0142] Unless otherwise stated, structures depicted herein are also
meant to include compounds which differ only in the presence of one
or more isotopically enriched atoms. For example, compounds having
the present structures except for the replacement of a hydrogen by
a deuterium or tritium, or the replacement of a carbon by .sup.13C-
or .sup.14C-enriched carbon are within the scope of this
disclosure.
[0143] The compounds of the present disclosure may also contain
unnatural proportions of atomic isotopes at one or more of atoms
that constitute such compounds. For example, the compounds may be
radiolabeled with radioactive isotopes, such as for example tritium
(.sup.3H), iodine-125 (.sup.125I) or carbon-14 (.sup.14C). All
isotopic variations of the compounds of the present disclosure,
whether radioactive or not, are encompassed within the scope of the
present disclosure.
[0144] As used herein the term "monomer" refers to a molecule that
can undergo polymerization, thereby contributing constitutional
units to the essential structure of a macromolecule or polymer.
[0145] A "polymer" is a molecule of high relative molecule mass,
the structure of which essentially comprises the multiple
repetition of unit derived from molecules of low relative molecular
mass, i.e., a monomer.
[0146] As used herein, an "oligomer" includes a few monomer units,
for example, in contrast to a polymer that potentially can comprise
an unlimited number of monomers. Dimers, trimers, and tetramers are
non-limiting examples of oligomers.
[0147] The compounds of the present disclosure may exist as salts.
The present disclosure includes such salts. Examples of applicable
salt forms include hydrochlorides, hydrobromides, sulfates,
methanesulfonates, nitrates, maleates, acetates, citrates,
fumarates, tartrates (e.g. (+)-tartrates, (-)-tartrates or mixtures
thereof including racemic mixtures, succinates, benzoates and salts
with amino acids such as glutamic acid. These salts may be prepared
by methods known to those skilled in art. Also included are base
addition salts such as sodium, potassium, calcium, ammonium,
organic amino, or magnesium salt, or a similar salt. When compounds
of the present disclosure contain relatively basic functionalities,
acid addition salts can be obtained by contacting the neutral form
of such compounds with a sufficient amount of the desired acid,
either neat or in a suitable inert solvent. Examples of acceptable
acid addition salts include those derived from inorganic acids like
hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic,
phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,
monohydrogensulfuric, hydriodic, or phosphorous acids and the like,
as well as the salts derived organic acids like acetic, propionic,
isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric,
lactic, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic,
citric, tartaric, methanesulfonic, and the like. Also included are
salts of amino acids such as arginate and the like, and salts of
organic acids like glucuronic or galactunoric acids and the like.
Certain specific compounds of the present disclosure contain both
basic and acidic functionalities that allow the compounds to be
converted into either base or acid addition salts.
[0148] The neutral forms of the compounds may be regenerated by
contacting the salt with a base or acid and isolating the parent
compound in the conventional manner. The parent form of the
compound differs from the various salt forms in certain physical
properties, such as solubility in polar solvents.
[0149] Certain compounds of the present disclosure can exist in
unsolvated forms as well as solvated forms, including hydrated
forms. In general, the solvated forms are equivalent to unsolvated
forms and are encompassed within the scope of the present
disclosure. Certain compounds of the present disclosure may exist
in multiple crystalline or amorphous forms. In general, all
physical forms are equivalent for the uses contemplated by the
present disclosure and are intended to be within the scope of the
present disclosure.
[0150] The term "pharmaceutically acceptable salts" is meant to
include salts of active compounds which are prepared with
relatively nontoxic acids or bases, depending on the particular
substituent moieties found on the compounds described herein. When
compounds of the present disclosure contain relatively acidic
functionalities, base addition salts can be obtained by contacting
the neutral form of such compounds with a sufficient amount of the
desired base, either neat or in a suitable inert solvent. Examples
of pharmaceutically acceptable base addition salts include sodium,
potassium, calcium, ammonium, organic amino, or magnesium salt, or
a similar salt. When compounds of the present disclosure contain
relatively basic functionalities, acid addition salts can be
obtained by contacting the neutral form of such compounds with a
sufficient amount of the desired acid, either neat or in a suitable
inert solvent. Examples of pharmaceutically acceptable acid
addition salts include those derived from inorganic acids like
hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic,
phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,
monohydrogensulfuric, hydriodic, or phosphorous acids and the like,
as well as the salts derived from relatively nontoxic organic acids
like acetic, propionic, isobutyric, maleic, malonic, benzoic,
succinic, suberic, fumaric, lactic, mandelic, phthalic,
benzenesulfonic, p-tolylsulfonic, citric, tartaric,
methanesulfonic, and the like. Also included are salts of amino
acids such as arginate and the like, and salts of organic acids
like glucuronic or galactunoric acids and the like {see, for
example, Berge et al, "Pharmaceutical Salts", Journal of
Pharmaceutical Science, 1977, 66, 1-19). Certain specific compounds
of the present disclosure contain both basic and acidic
functionalities that allow the compounds to be converted into
either base or acid addition salts.
[0151] In addition to salt forms, the present disclosure provides
compounds, which are in a prodrug form. Prodrugs of the compounds
described herein are those compounds that readily undergo chemical
changes under physiological conditions to provide the compounds of
the present disclosure. Additionally, prodrugs can be converted to
the compounds of the present disclosure by chemical or biochemical
methods in an ex vivo environment. For example, prodrugs can be
slowly converted to the compounds of the present disclosure when
placed in a transdermal patch reservoir with a suitable enzyme or
chemical reagent.
[0152] The term "protecting group" refers to chemical moieties that
block some or all reactive moieties of a compound and prevent such
moieties from participating in chemical reactions until the
protective group is removed, for example, those moieties listed and
described in T. W. Greene, P. G. M. Wuts, Protective Groups in
Organic Synthesis, 3rd ed. John Wiley & Sons (1999). It may be
advantageous, where different protecting groups are employed, that
each (different) protective group be removable by a different
means. Protective groups that are cleaved under totally disparate
reaction conditions allow differential removal of such protecting
groups. For example, protective groups can be removed by acid,
base, and hydrogenolysis. Groups such as trityl, dimethoxytrityl,
acetal and tert-butyldimethylsilyl are acid labile and may be used
to protect carboxy and hydroxy reactive moieties in the presence of
amino groups protected with Cbz groups, which are removable by
hydrogenolysis, and Fmoc groups, which are base labile. Carboxylic
acid and hydroxy reactive moieties may be blocked with base labile
groups such as, without limitation, methyl, ethyl, and acetyl in
the presence of amines blocked with acid labile groups such as
tert-butyl carbamate or with carbamates that are both acid and base
stable but hydrolytically removable.
[0153] Carboxylic acid and hydroxy reactive moieties may also be
blocked with hydrolytically removable protective groups such as the
benzyl group, while amine groups capable of hydrogen bonding with
acids may be blocked with base labile groups such as Fmoc.
Carboxylic acid reactive moieties may be blocked with
oxidatively-removable protective groups such as
2,4-dimethoxybenzyl, while co-existing amino groups may be blocked
with fluoride labile silyl carbamates.
[0154] Allyl blocking groups are useful in the presence of acid-
and base-protecting groups since the former are stable and can be
subsequently removed by metal or pi-acid catalysts. For example, an
allyl-blocked carboxylic acid can be deprotected with a
palladium(O)-- catalyzed reaction in the presence of acid labile
t-butyl carbamate or base-labile acetate amine protecting groups.
Yet another form of protecting group is a resin to which a compound
or intermediate may be attached. As long as the residue is attached
to the resin, that functional group is blocked and cannot react.
Once released from the resin, the functional group is available to
react.
[0155] Typical blocking/protecting groups include, but are not
limited to the following moieties:
##STR00009##
[0156] The subject treated by the presently disclosed methods in
their many embodiments is desirably a human subject, although it is
to be understood that the methods described herein are effective
with respect to all vertebrate species, which are intended to be
included in the term "subject." Accordingly, a "subject" can
include a human subject for medical purposes, such as for the
treatment of an existing condition or disease or the prophylactic
treatment for preventing the onset of a condition or disease, or an
animal subject for medical, veterinary purposes, or developmental
purposes. Suitable animal subjects include mammals including, but
not limited to, primates, e.g., humans, monkeys, apes, and the
like; bovines, e.g., cattle, oxen, and the like; ovines, e.g.,
sheep and the like; caprines, e.g., goats and the like; porcines,
e.g., pigs, hogs, and the like; equines, e.g., horses, donkeys,
zebras, and the like; felines, including wild and domestic cats;
canines, including dogs; lagomorphs, including rabbits, hares, and
the like; and rodents, including mice, rats, and the like. An
animal may be a transgenic animal. In some embodiments, the subject
is a human including, but not limited to, fetal, neonatal, infant,
juvenile, and adult subjects. Further, a "subject" can include a
patient afflicted with or suspected of being afflicted with a
condition or disease. Thus, the terms "subject" and "patient" are
used interchangeably herein.
[0157] In general, the "effective amount" of an active agent or
drug delivery device refers to the amount necessary to elicit the
desired biological response. As will be appreciated by those of
ordinary skill in this art, the effective amount of an agent or
device may vary depending on such factors as the desired biological
endpoint, the agent to be delivered, the composition of the
encapsulating matrix, the target tissue, and the like.
[0158] Following long-standing patent law convention, the terms
"a," "an," and "the" refer to "one or more" when used in this
application, including the claims. Thus, for example, reference to
"a subject" includes a plurality of subjects, unless the context
clearly is to the contrary (e.g., a plurality of subjects), and so
forth.
[0159] Throughout this specification and the claims, the terms
"comprise," "comprises," and "comprising" are used in a
non-exclusive sense, except where the context requires otherwise.
Likewise, the term "include" and its grammatical variants are
intended to be non-limiting, such that recitation of items in a
list is not to the exclusion of other like items that can be
substituted or added to the listed items.
[0160] For the purposes of this specification and appended claims,
unless otherwise indicated, all numbers expressing amounts, sizes,
dimensions, proportions, shapes, formulations, parameters,
percentages, parameters, quantities, characteristics, and other
numerical values used in the specification and claims, are to be
understood as being modified in all instances by the term "about"
even though the term "about" may not expressly appear with the
value, amount or range. Accordingly, unless indicated to the
contrary, the numerical parameters set forth in the following
specification and attached claims are not and need not be exact,
but may be approximate and/or larger or smaller as desired,
reflecting tolerances, conversion factors, rounding off,
measurement error and the like, and other factors known to those of
skill in the art depending on the desired properties sought to be
obtained by the presently disclosed subject matter. For example,
the term "about," when referring to a value can be meant to
encompass variations of, in some embodiments, .+-.100% in some
embodiments .+-.50%, in some embodiments .+-.20%, in some
embodiments .+-.10%, in some embodiments .+-.5%, in some
embodiments .+-.1%, in some embodiments .+-.0.5%, and in some
embodiments .+-.0.1% from the specified amount, as such variations
are appropriate to perform the disclosed methods or employ the
disclosed compositions.
[0161] Further, the term "about" when used in connection with one
or more numbers or numerical ranges, should be understood to refer
to all such numbers, including all numbers in a range and modifies
that range by extending the boundaries above and below the
numerical values set forth. The recitation of numerical ranges by
endpoints includes all numbers, e.g., whole integers, including
fractions thereof, subsumed within that range (for example, the
recitation of 1 to 5 includes 1, 2, 3, 4, and 5, as well as
fractions thereof, e.g., 1.5, 2.25, 3.75, 4.1, and the like) and
any range within that range.
EXAMPLES
[0162] The following Examples have been included to provide
guidance to one of ordinary skill in the art for practicing
representative embodiments of the presently disclosed subject
matter. In light of the present disclosure and the general level of
skill in the art, those of skill can appreciate that the following
Examples are intended to be exemplary only and that numerous
changes, modifications, and alterations can be employed without
departing from the scope of the presently disclosed subject matter.
The synthetic descriptions and specific examples that follow are
only intended for the purposes of illustration, and are not to be
construed as limiting in any manner to make compounds of the
disclosure by other methods.
Example 1
Methods
[0163] Murine Models of IBD: IL-10 Knockout (KO) and DSS-Induced
Colitis.
[0164] DSS-induced colitis model was generated as described
previously (Alex et al., 2009; Alex et al., 2010). IL-10-KO mice,
generated by gene-targeting, exhibited spontaneous colitis when
maintained in H. polyri-positive condition, is one of the most
widely used and most relevant murine models of colitis, was
described in details previously (Kuhn et al., 1993). The fact that
mutations in either IL-10 or IL-10 receptors in human lead to
spontaneous Crohn's disease at very early age (usually less than 1
year old) (Glocker et al., 2011), further emphasize the critical
role of IL-10 signaling pathway in its anti-inflammatory and
immune-modulatory effects in the gut and the homeostasis of healthy
gut.
[0165] Bioanalysis of 2-PMPA:
[0166] Blood and colonic mucosa were collected for drug PK
analysis. Plasma was generated from blood by centrifugation and all
samples were stored at -80.degree. C. until further analysis.
Concentrations of inhibitors in plasma and tissue were determined
via LC/MS/MS as described previously (Rais et al., 2013). Briefly,
2-PMPA was extracted from plasma and tissue by protein
precipitation with 5.times. methanol containing 2-(phosphonomethyl)
succinic acid (2-PMSA; 1 .mu.M) as an internal standard. The
samples were vortexed (30 s) and centrifuged (10,000 g for 10 min).
Supernatant was dried under a gentle stream of nitrogen at
45.degree. C. and the residue was reconstituted with 100 .mu.L of
acetonitrile and vortexed. 50 .mu.L of derivatizing agent
N-tert-Butyldimethysilyl-N-methyltrifluoro-acetamide (MTBSTFA) was
added to microcentrifuge tubes, vortexed, and heated at
approximately 60.degree. C. for 40 min. At the end of 40 min, the
derivatized samples were analyzed via LC/MS/MS. Chromatographic
analysis was performed using an Accela.TM. ultra high-performance
system consisting of an analytical pump, and an autosampler coupled
with TSQ Vantage mass spectrometer (Thermo Fisher Scientific Inc.,
Waltham Mass.). Separation of the analyte from potentially
interfering material was achieved at ambient temperature using
Waters X-terraR, RP18, 3.5 .mu.m, and (2.1.times.50 mm). The mobile
phase used was composed of 0.1% formic acid in acetonitrile and
0.1% formic acid in H.sub.2O with gradient elution, starting with
90% (organic) linearly increasing to 99% up to 2.5 min, maintaining
at 99% (2.5-4.0 min) and reequlibrating to 90% by 5 min. The total
run time for each analyte was 5.0 min. The [M+H]+ ion transitions
of derivatized 2-PMPA at m/z 683.0>551.4 and that of the
internal standard at m/z 669.0>537.2 were monitored with the
total run time of 5 min.
[0167] GCPII Ex Vivo Activity Assay:
[0168] On the day of the ex vivo experiment, the tissues were
weighed and immersed in 0.5 ml of ice-cold 50 mM Tris Buffer (pH
7.7 at RT). While on ice, each tissue was sonicated for 30-60
seconds (medium output, 60) via use of a Kontes Ultrasonic Cell
Disrupter. After a 2 min spin at 13,000 rpm, supernatants were
analyzed for protein content (Dc Protein Assay Kit; Bio Rad).
NAAG-hydrolyzing activity measurements were carried out, based on
published procedures Robinson et al., 1987; Rojas et al., 2002).
Briefly, the reaction mixture (total volume of 50 .mu.L) contained
NAA[3H]G (70 nM, 50 Ci/mmol) and tissue lysates (7 concentrations
each, tested in quadruplicate), in Tris-HCl (pH 7.4, 40 mM)
containing 1 mM CoCl.sub.2. Assay standards were comprised of human
recombinant GCPII enzyme (40-200 pM, final). The reaction was
carried out at 37.degree. C. for 40-45 min and stopped with
ice-cold sodium phosphate buffer (pH 7.4, 0.1 M, 50 .mu.L). Blanks
were obtained by incubating the reaction mixture in the absence of
enzyme source. A 90 .mu.L aliquot from each terminated reaction was
transferred to a well in a 96-well spin column containing AG1X8
ion-exchange resin; the plate was centrifuged at 1500 rpm for 5 min
using a Beckman GS-6R centrifuge equipped with a PTS-2000 rotor.
NAA[3H]G bound to the resin and [3H]-G eluted in the flow through.
Columns were then washed twice with formate (1 M, 90 .mu.L) to
ensure complete elution of [3H]-G. The flow through and the washes
were collected in a deep 96-well block; from each well with a total
volume of 270 .mu.L, a 200 .mu.L aliquot was transferred to its
respective well in a solid scintillator-coated 96-well plate
(Packard) and dried to completion. The radioactivity corresponding
to [3H]-G was determined with a scintillation counter (Topcount
NXT, Packard, counting efficiency 80%). Enzymatic activity (fmol/mg
total protein/hour) determinations were based on slopes of lines
obtained from CPM.fwdarw.DPM.fwdarw.molar conversion per unit time
vs protein concentrations. Results were generated via use of
Microsoft Office Excel 2007 and GraphPad Prism 5 programs.
Example 2
PSMA Expression and Enzymatic Activity is Selectively Elevated in
Patient Samples with IBD
[0169] Previously gene-profiling and immuno-histological analyses
(FIG. 1A and FIG. 1B) showed that PSMA is significantly upregulated
in the intestinal mucosa of patients with Crohn's disease (Zhang et
al., 2012). To further determine the relevance of PSMA to IBD, PSMA
functional enzymatic activity was examined in normal and diseased
mucosa of 32 surgical intestinal specimens from 20 subjects (FIG.
2A and FIG. 2B), including healthy controls, patients with IBD, and
non-IBD controls (diverticulitis). A very significant and robust
300-1,000% increase in PSMA activity was found in the intestinal
mucosa with active IBD when compared to that in an uninvolved area
of the same patients, or the intestine from healthy and non-IBD
controls. These data suggest a clear positive association between
activation of PSMA and IBD.
Example 3
Preclinical Efficacy: 2-PMPA, a Potent and Selective PSMA
Inhibitor(PSMAi), Shows Profound Efficacy in Two Major Animal
Models of IBD
[0170] To investigate whether PSMA can be a suitable novel
therapeutic target for clinical intervention against IBD, the
effect of PSMA prototype inhibitors on two most widely used murine
models of IBD, DSS-induced colitis, and IL-10 knockout (IL-10 KO)
mice (a genetic model that develops spontaneous colitis), was
tested. In both models, PSMA inhibitor treatment dramatically
ameliorated symptoms. In the DSS colitis model, PSMA inhibition
significantly reduced the disease activity index (FIG. 3).
Moreover, the PSMA activity in the colonic and cecal mucosa of
DSS-treated mice was potently inhibited by 2-PMPA, indicating
target engagement (FIG. 4). 2-PMPA also significantly reduced
spleen inflammation (FIG. 5) on mice with DSS-colitis, suggesting
its immune-suppressive effect on systemic inflammation, in addition
to gut inflammation.
[0171] The efficacy of 2-PMPA in treatment of spontaneous colitis
in IL-10 KO mice was also remarkable. 2-PMPA significantly reduced
the disease severity, including macroscopic disease, colonic
hypotrophy, and provided better stool consistency (FIGS. 6A-6B).
More interestingly, a complete retraction of prolapse was observed
in 2 of the 20 mice (10%) treated with the inhibitor (FIG. 6D), a
phenomenon that has never been seen in more than 800 IL-10 KO mice
used in previous efforts. The improvement of these
prolapse-retracting mice was unequivocally obvious in that their
body weight increased dramatically when compared to that of
untreated control IL-10 KO mice (FIG. 6C). In conclusion, using
three major animal models of IBD, the significance of PSMA as a
novel therapeutic target for treatment of IBD was demonstrated.
Example 4
Two Structurally Distinct PSMA Inhibitors (E2072 and 2-PMPA) Show
Similar Efficacy in IBD Models
[0172] E2072 ((3-2-Mercaptoethyl)biphenyl-2,3-dicarboxylic acid,
another PSMA inhibitor that is structurally distinct from 2-PMPA,
exhibited similar ameliorating effects on DSS-induced colitis (FIG.
7).
Example 5
Oral Administration of TRIS POC 2-PMPA (Novel Prodrug of 2-PMPA)
Exhibits Comparable Concentrations in Plasma and Colon to Those of
2-PMPA Given i.p
[0173] 2-PMPA demonstrated excellent efficacy following i.p.
administration at 100 mg/kg in both the DSS and IL 10 knock out
model (FIGS. 4 and 6). An orally bioavailable prodrug of 2-PMPA
that enabled approximately 20 fold enhancement in permeability
(FIG. 8A) in mice has now been identified. More importantly, the
prodrug exhibits similar concentrations to 100 mg/kg i.p.
administration when dosed p.o. FIG. 8A shows direct comparison of
prodrug (30 mg/kg equiv) to 2-PMPA (100 mg/kg i.p.) in plasma and
colon samples collected after 8 days of daily dosing in DSS mice.
On day 8 the animals were sacrificed 2 h after dosing. The prodrug
at a lower dose demonstrated comparable concentrations in both
plasma and colon (approximately 15 .mu.M at 30 mg/kg) compared to
2-PMPA (approximately 25 .mu.M at 100 mg/kg).
Example 6
Oral Administration of TRIS POC 2-PMPA
[0174] Oral administration of TRIS POC 2-PMPA exhibited similar
ameliorating effects on DSS-induced colitis, although a larger
number of mice are needed in each experimental group to achieve
statistical significance. As shown in FIG. 9, colon shortening (a
signature of DSS-colitis) of DSS-colitis mice was reversed after
oral administration of prodrug Tris POC 2-PMPA.
[0175] The presently disclosed genomic, clinical, and
pharmacological data implicate PSMA in the etiology of inflammatory
bowel disease (IBD). The data illustrate that PSMA enzymatic
activity is consistently and robustly activated with the human
clinical disease and that pharmacological inhibition of PSMA using
multiple structurally distinct inhibitors ameliorate IBD
symptoms.
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[0176] All publications, patent applications, patents, and other
references mentioned in the specification are indicative of the
level of those skilled in the art to which the presently disclosed
subject matter pertains. All publications, patent applications,
patents, and other references (e.g., websites, databases, etc.)
mentioned in the specification are herein incorporated by reference
in their entirety to the same extent as if each individual
publication, patent application, patent, and other reference was
specifically and individually indicated to be incorporated by
reference. It will be understood that, although a number of patent
applications, patents, and other references are referred to herein,
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are used herein unless indicated otherwise. Standard abbreviations
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[0209] Although the foregoing subject matter has been described in
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* * * * *
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